Mitochondria tethered Pink1 overexpression restores tamm41 deficiency induced heart valve abnormalities. a Representative images show the recovering effect of opa3-pink1 overexpression on 56 hpf tamm41−/− heart valve via DR-GRASP staining. Scale bar: 20 μm. bOpa3-pink1 injection at one-cell stage embryos restored heart valve markers, including bmp4 and notch1b, in tamm41−/− embryos. c, d Reduced mitochondrial membrane protein levels (fourth lane) in tamm41−/− embryos with Opa3-pink1 mRNA injection. e, f OPA3-PINK1 overexpression displayed reduced mitochondrial membrane proteins in si-TAMM41 AC16 cells with CCCP addition (fourth panel). g Representative images (n = 50 per group) revealed mito-tethered PINK1 (OPA3-PINK1) overexpression restored CCCP triggered mitophagy activity in the TAMM41 deficient AC16 cell line. Scale bar: 5 μm. h Representative images (n = 50 cells per group) show the enforced OPA3-PINK1 restored HA-PARK2 translocation onto mitochondria (indicated by white arrows) in the TAMM41 deficient AC16 cell line with CCCP treatment. Scale bar: 20 μm. i, j (i) and (j) are quantitative data for (g) and (h) respectively. Black horizontal lines indicate mean ± SD. Means ± SD are shown for three independent experiments. n.s, not significant, *P < 0.05; **P < 0.01; ***P < 0.001 (Student’s t-test)
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